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Determination of glycosidase activity in porcine oviductal fluid at the different phases of the estrous cycle

¹ Department of Physiology, Faculty of Veterinary Science, University of Murcia, 30071 Murcia, Spain² Department of Biology, Faculty of Basic Sciences, University of Pamplona, 2-102 El Carmen, Santander del Norte, Pamplona, Colombia³ Department of Cell Biology and Histology, Faculty of Medicine, University of Murcia, 30071 Murcia, Spain

Correspondence should be addressed to R Romar; Email: rromar{at}um.es

Sperm–oocyte binding and gamete–oviductal epithelium interactions are carbohydrate-mediated events occurring in the oviductal fluid (OF). Thus, knowledge about the activities of glycosidases (enzymes catalyzing hydrolytic cleavage of terminal sugar residues) in this milieu would help us understand the molecular mechanisms involved in these events. This work was carried out to investigate the glycosidase activity, protein content, and volume of OF collected from gilts and sows. Oviducts were classified into four phases of the estrous cycle (early follicular, late follicular, early luteal, and late luteal) based on the appearance of the ovaries. OF was aspirated, centrifuged, measured for volume, and frozen until assay. Substrates conjugated to 4-methylumbelliferyl were used to screen the activities of seven different glycosidases at physiological pH (7.2). -L-Fucosidase and β-N-acetyl-glucosaminidase activities increased at the late follicular phase to decrease after ovulation. β-D-Galactosidase, -D-mannosidase, and β-N-acetyl-galactosaminidase showed higher activities at the early follicular phase, which decreased after ovulation. N-Acetyl-neuraminidase and -D-galactosidase did not show activity at any phase of estrous cycle neither in sows nor in gilts at pH 7.2, although it did at acidic pH (4.4) in the follicular and luteal phase samples. Total protein also changed during the cycle showing the maximum secretion at the late follicular phase (2118.6±200.7 µg/oviduct). The highest volumes of OF were collected from the oviducts at the late follicular phase (50.7±1.3 µl/oviduct). These results indicate that OF from sows and gilts shows glycosidase activity varying throughout the estrous cycle suggesting a role of these enzymes in carbohydrate-mediated events.

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				R. E Lloyd, R. Romar, C. Matas, A. Gutierrez-Adan, W. V Holt, and P. Coy Effects of oviductal fluid on the development, quality, and gene expression of porcine blastocysts produced in vitro Reproduction, April 1, 2009; 137(4): 679 - 687. [Abstract] [Full Text] [PDF]